Canola cultivar g069467

ABSTRACT

The present invention relates to a new and distinctive canola, designated G069467. Also included are seeds of canola G069467, to the plants, or plant parts, of canola G069467 and to methods for producing a canola plant produced by crossing the canola G069467 with itself or another canola genotype, and the creation of variants by mutagenesis or transformation of canola G069467.

FIELD OF THE INVENTION

This invention relates to a canola inbred cultivar designated G069467that includes plants, DNA, and seeds of canola G069467. Methods forproducing canola plants, such as canola plant varieties, hybrid canolaplants, or other canola plants, as by crossing canola G069467 withitself or any different canola plant are an integral part of thisinvention as are the resultant canola plants including the plant partsand seeds. This invention further relates to methods for producingG069467-derived canola plants, to methods for producing male sterileG069467 canola plants, e.g., cytoplasmic male sterile G069467 canolaplants and to methods for regenerating such plants from tissue culturesof regenerable cells as well as the plants obtained there from. Methodsfor producing a canola plant from G069467 containing in its geneticmaterial one or more transgenes and to the transgenic canola plantsproduced by that method are also part of this invention.

BACKGROUND OF THE INVENTION

“Canola”, refers to a particular class of rapeseed (Brassica napusoleifera annua) having: (i) a seed oil that contains less than 2% erucicacid, and (ii) an oil-free meal that contains less than 30 micromolesaliphatic glucosinolates per gram of meal. Canola seed is pressed forcooking oil and the residual meal is used as an organic fertilizer andas a high-protein animal feed supplement. Industrial uses of canolainclude biodiesel and plastic feedstocks.

Farmers in Canada began producing canola oil in 1968. Early canolacultivars were known as single zero cultivars because their oilcontained 5% or less erucic acid, but glucosinolates were high. In 1974,the first licensed double zero cultivars (low erucic acid and lowglucosinolates) were grown. Today all canola cultivars are double zerocultivars. The Canadian Health and Welfare Department recommendedconversion to the production of low erucic acid varieties of rapeseed.Industry responded with a voluntary agreement to limit erucic acidcontent to less than 5% in food products, effective Dec. 1, 1973. In1985, the U.S. Food and Drug Administration granted canola oil GRAS(Generally Recognized as Safe) status for use in human foods.

Because canola oil is perceived to be “healthy”, its use is risingsteadily both as an oil for cooking and as an ingredient in processedfoods. The consumption of canola oil is expected to surpass corn andcottonseed oils, becoming second only to soybean oil. It is low insaturated fatty acids and high in monounsaturated fatty acids,containing a high level of oleic acid. Many people prefer the lightcolor and mild taste of canola oil over olive oil, the other readilyavailable oil high in monounsaturates.

Canola is an important and valuable field crop. The goal of a canolabreeder is to develop new, unique, and superior canola cultivars andhybrids having improved combinations of desirable traits and therefore,increased economic value. Improved performance is manifested in manyways. Higher yields of canola plants contribute to higher seedproduction per acre, a more profitable agriculture and a lower cost ofproducts for the consumer. Improved plant health increases the yield andquality of the plant and reduces the need for application of protectivechemicals. Adapting canola plants to a wider range of production areasachieves improved yield and vegetative growth. Improved plant uniformityenhances the farmer's ability to mechanically harvest canola. Improvednutritional quality increases its value in food and feed.

Canola is a dicot plant with perfect flowers, i.e., canola has male,pollen-producing organs and separate female, pollen receiving organs onthe same flower. Canola flowers are radial with four sepals alternatingwith four petals forming the typical cross pattern from which theCruciferae family derives its name. In addition, canola flowers consistof two short lateral stamens, four longer median stamens and a stigma.Pollination occurs with the opening of the anthers and shedding ofpollen on the stigma or with the deposit of pollen on the stigma byinsects. Canola flowers are mainly self-pollinating, althoughoutcrossing occurs when pollen is transferred from the anthers to thestigmas by wind or bees or other insects. After fertilization, which isusually complete within 24 hours of pollination, the syncarpous ovaryelongates to form a silique (pod). Because each pod may contain 25 ormore seeds and each plant produces many pods, the multiplication rateper generation usually exceeds 1,000 to 1, thereby accelerating thebreeding and evaluation process.

The development of new cultivars in a canola plant breeding programinvolves numerous steps, including: (1) selection of parent canolaplants (germplasm) for the initial breeding crosses; (2) producing andselecting inbred breeding lines and cultivars by either thedoubled-haploid method or repeated generations of selfing individualplants, which eventually breed true; (3) producing and selecting hybridcultivars by crossing a selected inbred male-sterile line with anunrelated inbred restorer line to produce the F₁ hybrid progeny havingrestored vigor; and (4) thoroughly testing these advanced inbreds andhybrids compared to appropriate standards for three or more years inenvironments representative of the commercial target area(s). The bestinbred and hybrid experimental cultivars then become candidates for newcommercial cultivars. Those lines still deficient in a few traits may beused as parental lines to produce new populations for further selection.

Development and selection of new canola parental lines, the crossing ofthese parental lines, and selection of superior hybrid progeny are vitalto maintaining a canola breeding program. The F₁-hybrid canola seed isproduced by manual crosses between selected male-fertile parents or byusing male-sterility systems. These hybrids are selected for certainsingle-gene traits such as pod color, flower color, pubescence color orherbicide resistance, which can indicate that the seed is truly a hybridfrom the intended cross. Additional data on parental lines, as well asthe phenotype of the hybrid, influence the breeder's decision whether tocontinue with the specific hybrid cross.

The method of doubled-haploid breeding consists of donor selection,microspore culture and chromosome doubling, embryo cold stress, plantletregeneration, ploidy analysis, and self-pollination to produce seed ofdoubled-haploid lines. The advantage of the doubled-haploid method isthat the time to develop a new completely homozygous and homogeneouscultivar can be reduced by 3 years compared to the conventionalinbreeding method of multiple generations of self pollination.

When two different, unrelated canola parent cultivars are crossed toproduce an F₁ hybrid, one parent cultivar is designated as the male, orpollen parent, and the other parent cultivar is designated as thefemale, or seed parent. Because canola plants are capable ofself-pollination, hybrid seed production requires elimination of orinactivation of pollen produced by the female parent. Different optionsexist for controlling male fertility in canola plants such as physicalemasculation, application of gametocides, and cytoplasmic male sterility(CMS).

Hybrid canola seed can be produced on a commercial scale by means of asystem whereby the female parent has an allele in the mitochondrialgenome for cytoplasmic male sterility and the male parent has an allelein the nuclear genome for fertility restoration (Rf). Cytoplasmic malesterility prevents the production of functional pollen, therebypreventing self pollination of the female parent. Pollen from the maleparent planted in close proximity to the female parent is then able tofreely cross pollinate the female parent to produce hybrid seed. Thefertility-restoration allele contributed by the male parent to the seedembryo enables the hybrid crop plants to be male fertile. The resultinghybrid canola crop, which is fully fertile, may then demonstrateheterosis (increased vigor) to produce grain yields potentially greaterthan that of inbred cultivars.

A cytoplasmic male-sterile inbred (A) line is genetically maintained andincreased in a breeding and hybrid-production program by growing it inisolation with a male-fertile maintainer (B) line that is normal (N) forcytoplasmic fertility and is homozygous recessive at the nuclearmale-fertility restoration locus (N rfrf). All seed harvested from the Aline is then male sterile (S rfrf) and all seed harvested off the B lineis male fertile (N rfrf). The A line is then maintained, increased, andused as the female parent for hybrid seed production in combination withan unrelated male parent that has the dominant allele (Rf) for malefertility restoration.

One example of a CMS system in canola hybrid production and breeding isthe Ogura (Ogu) cytoplasm and its specific nuclear fertility-restorationgene, Rfo—a system discovered in radish (Raphanus sativus) andtransferred to Brassica napus after protoplast fusion. The system waslater improved by breeding to lower the glucosinolate content for hybridcanola.

These processes, which lead to the final step of marketing anddistribution of a cultivar, usually take from 8 to 12 years from thetime the parental cross is made. Therefore, development of new canolainbred and hybrid cultivars is a slow, costly process that requires theresources and expertise of plant breeders and numerous otherspecialists.

It is nearly impossible for two canola breeders to independently developgenetically-identical canola inbreds or hybrids expressing all the sametrait characteristics. The cultivars that are developed cannot bepredicted because the breeder's selection occurs in unique environments,with no control over meiotic genetic recombination (using conventionalbreeding procedures), and with millions of different possible geneticcombinations possible. A breeder of ordinary skill in the art cannotpredict the final resulting lines he/she develops, except possibly in avery gross and general fashion. The same breeder cannot produce the samecultivar twice by using the exact same original parents and the sameselection techniques.

Canola cultivars and other sources of canola germplasm are thefoundation material for all canola breeding programs. Despite theexistence and availability of numerous canola cultivars and other sourcegermplasm, a need still exists for the development of improved germplasmto improve and maximize yield and quality.

The foregoing examples of the related art and limitations relatedtherewith are intended to be illustrative and not exclusive. Otherlimitations of the related art will become apparent to those of skill inthe art upon a reading of the specification.

SUMMARY OF THE INVENTION

The following embodiments and aspects thereof are described inconjunction with systems, tools and methods that are meant to beexemplary and illustrative, not limiting in scope.

According to the invention, there is provided a novel canola cultivardesignated G069467. This invention thus relates to the seeds of canolaG069467, to the plants, or plant parts, of canola G069467 and to methodsfor producing a canola plant produced by crossing the canola G069467with itself or another canola cultivar, and the creation of variants bymutagenesis or transformation of canola G069467.

Thus, any such methods using the canola G069467 are part of thisinvention: selfing, backcrossing, hybrid production, crossing topopulations, and the like. All plants produced using canola G069467 as aparent, are within the scope of this invention. Advantageously, thecanola G069467 could be used in crosses with other, different, canolaplants to produce first generation (F₁) canola hybrid seeds and plantswith superior characteristics.

In another aspect, the present invention provides for single or multiplegene-converted plants of canola G069467. The transferred gene(s) maypreferably have dominant or recessive allele(s). Preferably, thetransferred gene(s) will confer such traits as herbicide resistance,insect resistance; resistance to bacterial, fungal, or viral disease;male fertility, male sterility, enhanced nutritional quality, orindustrial usage. The gene may be a naturally occurring canola gene or atransgene introduced through genetic engineering techniques.

In another aspect, the present invention provides regenerable cells foruse in tissue culture of canola G069467. The tissue culture willpreferably be capable of regenerating plants having the physiologicaland morphological characteristics of the foregoing canola plant, and ofregenerating plants having substantially the same genotype as theforegoing canola plant. Preferably, the regenerable cells in such tissuecultures will be embryos, protoplasts, meristematic cells, callus,pollen, leaves, anthers, roots, root tips, flowers, seeds, pods orstems. Still further, the present invention provides canola plantsregenerated from the tissue cultures of the invention.

In another aspect, the present invention provides a method ofintroducing a desired trait into canola G069467 wherein the methodcomprises: crossing a G069467 plant with a plant of a different canolagenotype that comprises a desired trait to produce F₁ progeny plants,wherein the desired trait is selected from the group consisting of malesterility, herbicide resistance, insect resistance, and resistance tobacterial disease, fungal disease or viral disease; selecting one ormore progeny plants that have the desired trait to produce selectedprogeny plants; crossing the selected progeny plants with the G069467plants to produce backcross progeny plants; selecting for backcrossprogeny plants that have the desired trait and physiological andmorphological characteristics of canola G069467 to produce selectedbackcross progeny plants; and repeating these steps to produce selectedfirst or higher backcross progeny plants that comprise the desired traitand all of the physiological and morphological characteristics of canolaG069467. Included in this aspect of the invention is the plant producedby the method wherein the plant has the desired trait and all of thephysiological and morphological characteristics of canola G069467.

In addition to the exemplary aspects and embodiments described above,further aspects and embodiments will become apparent by study of thefollowing descriptions.

DETAILED DESCRIPTION OF THE INVENTION

In the description and examples that follow, a number of terms are used.To provide a clear and consistent understanding of the specification andclaims, including the scope to be given such terms, the followingdefinitions are provided.

Definitions of Plant Characteristics

Allele: Any of one or more alternative forms of a gene, all of whichrelate to the same protein, trait, or characteristic. In a diploid cellor organism, the alleles of a given gene occupy the corresponding locuson a pair of homologous chromosomes.

Anther Arrangement: The orientation of the anthers in fully openedflowers rated as introse (facing inward toward pistil), erect (neitherinward not outward), or extrose (facing outward away from pistil).

Anther Dotting: The presence or absence of colored spots on the tips ofanthers and if present, the percentage of anthers with dotting in newlyopened flowers.

Anther Fertility: The amount of pollen produced on the anthers of aflower; rated as sterile (such as in female parents used for hybrid seedproduction) to fertile (all anthers shedding).

AOM hours: A measure of the oxidative stability of an oil usingcurrently accepted Official Methods of the American Oil Chemists'Society (eg, AOCS 12b 92).

Backcrossing: A process in which a breeder repeatedly crosses hybridprogeny back to one of the parents; for example, a first generationhybrid F1 crossed back to one of the parental genotypes of the F1hybrid.

Blackleg Resistance: Resistance of a plant to two fungi (Leptosphaeriamaculans and Leptosphaeria biglobosa) that cause a disease commonlycalled blackleg; visually rated relative to a highly susceptible checksuch as the cultivar ‘Westar’ as Resistant (<30% of Westar), ModeratelyResistant (30% to 49% of Westar), Moderately Susceptible (50% to 69% ofWestar), Susceptible (70% to 100% of Westar), or Highly Susceptible(>100% of Westar).

Chlorophyll Content: The chlorophyll content of grain; measured usingnear-infrared (NIR) spectroscopy or other methods recommended by theWestern Canada Canola/Rapeseed Recommending Committee (WCC/RRC); ratedas low (<8 ppm), medium (8 to 15 ppm), and high (>15 ppm).

Cotyledon Width: The maximum width of a fully developed cotyledon (firstphotosynthetic leaves) measured around 2-3 weeks after emergence whenthe plant is at the 2-leaf to 3-leaf stage of development and is ratedas narrow, medium, or wide.

Cultivar: A plant genotype that has been intentionally bred or selectedto be genetically distinct, uniform, and stable, and is maintainedthrough cultivation or other propagation.

Cytoplasmic Male Sterility (CMS): A system by which a plant is unable toproduce functional pollen as a result of a genetic abnormality at alocus in its mitochondrial genome. Male fertility may be restored whenthis male-sterile cytoplasm is combined by crossing to a genotype with aparticular complementary allele at a locus in the nuclear genome thatsuppresses this cytoplasmic dysfunction.

Days to Flowering: The number of days from planting to the stage when50% of the plants show one or more open flowers.

Days to Maturity: The maturity of a variety measured as the number ofdays between planting and physiological maturity, i.e., the date whenpods ⅓rd up the main raceme have 60% of the seeds expressing a colorchange from green to brown or black.

Doubled Haploid (Dihaploid): A genotype formed when haploid cellsundergo chromosome doubling. Used in plant breeding to developcompletely homozygous lines faster than can be done by multiplegenerations of inbreeding.

Early-Season Vigor (ESV): Assessed at the 4- to 5-leaf stage taking intoaccount plant stand (or emergence) and ground cover of the desired crop;rated on a 1 to 5 scale where 1 is strong vigor and 5 is weak vigor.

Eicosenoic Acid: The total of eicosenoic acid (C20:1) in an oil sampleobtained from unprocessed seed; measured as a percentage of total oil.

Elite canola: A canola cultivar that has been stabilized for certaincommercially important agronomic traits wherein the relative value of atrait is about 100% or greater relative to the check cultivars growingin the same field location at the same time and under the sameconditions. In one embodiment, “elite canola” means a canola cultivarstabilized for yield of 110% or greater relative to the yield of checkcultivars growing in the same field location at the same time and underthe same conditions. In another embodiment, “elite canola” means acanola cultivar stabilized for yield of 115% or greater relative to theyield of check cultivars growing in the same field location at the sametime and under the same conditions.

Embryo: The genetic precursor tissue to the plant contained within andgerminating from a mature seed.

Erucic Acid: The total erucic acid (C22:1) in an oil sample obtainedfrom unprocessed seed; measured as a percentage of total oil.

FAME analysis: Quantification of fatty acid methyl esters (FAME)resulting from methylation to separate individual fatty acids from theglycerol backbone in triglycerides.

Fatty Acids: Various sizes of saturated and unsaturated hydrocarbonchains that are found in oil; of which individual sizes can be measuredusing gas chromatography or Near Infrared Resonance (NIR) spectroscopyas percentages of the total oil extracted from unprocessed seed.

Fertility Restoration: A system by which a dominantfertility-restoration allele (Rf) at a locus in the nuclear genomerestores male fertility to a hybrid progeny plant resulting from a crossof this allele to a parent having a compatible cytoplasmicmale-sterility (CMS) system.

Flower Bud Location: The location of unopened flower buds relative toadjacent opened flowers rated as above or below the opened flower. Theunopened buds are positioned above the most recently opened flowers inBrassica napus and positioned below the most recently opened flower budsin Brassica rapa.

Fusarium-Wilt Resistance: Resistance to two soil-borne fungi (Fusariumavenaceum and Fusarium oxysporum) that cause a disease commonly calledFusarium wilt; visually rated relative to a highly susceptible check asResistant (<30% of check), Moderately Resistant (30% to 49% of check),Moderately Susceptible (50% to 69% of check), Susceptible (70% to 100%of check), or Highly Susceptible (>100% of check).

Genotype: Refers to either the complete genetic composition or aspecific portion of the genetic composition of an organism.

Glucosinolate Content: The total aliphatic glucosinolates expressed asmicromoles per gram of whole seed. Glucosinolates can be measured byreversed-phase high-performance liquid chromatography (HPLC) (AOCSOfficial Method Ak 1-92), by capillary gas chromatography of thetrimethylsityl derivatives of extracted and purifieddesulfoglucosinolates (Procedures of the Western Canada Canola/RapeseedRecommending Committee), or by near-infrared (NIR) spectroscopy (AOCSOfficial Method Am 1-92).

Glyphosate Herbicide Resistance: Resistance of a plant to the action ofglyphosate; conferred in crops by genetic transformation of the cropplant using a 5-enol-pyruvylshikimate-3-phosphate synthase (EPSPS) genethat is insensitive to the effect of glyphosate, or a bacterialglyphosate oxidoreductase (GOX) gene that cleaves the nitrogen-carbonbond in glyphosate yielding aminomethylphosphonic acid.

Herbicide Resistance: When a plant has negligible effect from contactwith an herbicide because the plant does not take up the herbicide orsequesters the herbicide in a manner that renders it harmless.

Herbicide Tolerance: When a plant has negligible effect from contactwith an herbicide because the plant metabolically detoxifies theherbicide.

Hybrid: A cultivar or plant-breeding progeny based upon the controlledcross-pollination between or among distinct parent lines, so that theresulting seed inherits its genetic composition from those parent lines.Seed for a particular hybrid can be repeatedly and predictably producedwhen repeatedly making controlled cross-pollinations from the samestable female and male parent genotypes.

Imidazolinone (Imi) Tolerance: Tolerance of a plant to the action ofimidazolinone; conferred by one or more genes that alter acetolactatesynthase (ALS), also known as acetohydroxy acid synthase (AHAS), to beinsensitive to imidazolinone thereby preventing injury when exposed tothis class of herbicides.

Inbred: A relatively stable plant genotype resulting from doubledhaploids, successive generations of controlled self-pollination,successive generations of conrolled backcrossing to a recurrent parent,or other method to develop homozygosity.

Leaf Attachment to Stem: The degree to which the base of the leaf bladeof the supper stem leaf clasps the stem rated as complete clasping(Brassica rapa), partial clasping (Brassica napus), or non-clasping(mustard species).

Leaf Color: Leaf blade color recorded at the 5-leaf stage and rated aslight green, medium green, dark green, or blue green.

Leaf Glaucosity: Leaf waxiness determined by rubbing the leaf surface;rated as absent or present and, if present, as very weak, weak, medium,strong, or very strong.

Leaf Length: The length of the leaf from the tip to the base of thepetiole; visually determined to be short, medium, or long.

Leaf Lobe: A leaf lobe exists when the leaf tissue is indented in twoplaces to at least half the distance to the midrib. The upper area ofthe leaf is counted as a lobe (terminal lobe).

Leaf Lobe Development: Lobe development on a fully developed upper-stemleaf before flowering when at least 6 leaves of the plant are completelydeveloped; visually rated as absent or present and, if present, as veryweak, weak, medium, strong, or very strong.

Leaf Lobe Number: Count of the leaf lobes on a fully developedupper-stem leaf before flowering when at least 6 leaves of the plant arecompletely developed.

Leaf Lobe Shape: Shape of the leaf lobes; visually determined to beacute or rounded.

Leaf Margin Indentation: The serration along the margin of a fullydeveloped upper stem leaf measured before flowering when at least 6leaves of the plant are completely developed; rated as absent or presentand, if present, as very weak, weak, medium, strong, or very strong.

Leaf Margin Shape: Shape of the leaf margin on the upper third of thelargest leaf; visually determined to be undulating, rounded, or sharp.

Leaf Pubescence: Degree of hairiness on a leaf surface; rated asglabrous (smooth/not hairy, Brassica napus) or pubescent (hairy,Brassica rapa).

Leaf Shape: Shape of the leaf; visually determined as narrow elliptic(width/length <0.67), wide elliptic (width/length =0.67 to 0.79), ororbicular (width/length >0.79).

Leaf Surface Texture: Degree of leaf-surface wrinkling when at least 6leaves of the plant are completed developed; rated as smooth or rough.

Leaf Texture: A description of the texture of the leaf surface; visuallydetermined to be smooth or rough.

Leaf Type: Leaf visually rated as petiolate or lyrate.

Leaf Waxiness: A description of the waxiness on the surface of the leaf;visually determined to be absent, weak, medium, strong, or very strong.

Leaf Width: The width of the leaf between its widest points on bothedges on opposite sides of the midrib; visually determined to be narrow,medium, or wide.

Linoleic Acid: The total linoleic acid (C18:2) in an oil sample obtainedfrom unprocessed seed; measured as a percentage of total oil.

Linolenic Acid: The total linolenic acid (C18:3) in an oil sampleobtained from unprocessed seed; measured as a percentage of total oil.

Lodging Resistance: A scale for the physical orientation of plants afterexposure to adverse environmental conditions; generally measured byobserving the angle of the stem to the ground on a 1 to 5 scale where1=fully erect, 3=45° angle between stem and the ground, and 5=flat.

Lyrate Leaf: Leaf with laminar tissue continuing along the whole lengthof the petiole to the auricles.

Near Infrared (NIR) Spectroscopy: A spectroscopic method that uses thenear-infrared region of the electromagnetic spectrum (from about 800 nmto 2500 nm).

Oil Content: The amount of oil in unprocessed grain as a percentage ofthe whole dried seed; determined by pulsed nuclear magnetic resonance(NMR) (AOCS Official Method Ak 4-95) or near-infrared (NIR) spectroscopy(AOCS Official Method Am 1-92).

Oleic Acid: The total oleic acid (C18:1) in an oil sample obtained fromunprocessed seed; measured as a percentage of total oil.

Palmitic Acid: The total palmitic acid (C16:0) in an oil sample obtainedfrom unprocessed seed; measured as a percentage of total oil.

Petal Color: Flower petal color on open exposed petals; rated the firstday of flowering as white, light yellow, medium yellow, dark yellow,orange, or other.

Petal Length: Length of petals; visually determined to be short, medium,or long.

Petal Width: Width of petals; visually determined to be narrow, medium,or wide.

Petal Spacing: The proximity of petals to each other on a fully openedflower; visually determined to be open (large space between petals), nottouching (small space between petals), touching, slight overlap, orstrong overlap.

Petiolate Leaf: Leaf with a distinct and mostly naked petiole; somediscrete petiolar bracts may be present.

Petiole Length: Length of the petiole between where it is attached tothe stem and where, at the highest point on the leaf, leaf tissue eitherjoins the midrib or is within 4mm of the midrib; visually determined tobe short, medium, or long.

Plant Growth Habit: The angle of the outermost fully-expanded leafpetioles relative to the soil surface measured at end of flowering;rated as erect (>85°), semi-erect (approximately 65°), semi-prostrate(approximately 45°), or prostrate (<30°).

Plant Height: The height of a plant at the end of flowering from theground to the top of floral branches that are extended upright (ie, notlodged); visually determined to be short, medium, or tall.

Pod (silique) Angle: The orientation of the pods along the racemes(flowering stems); visually determined to be erect (pods angled close toracemes), semi-erect (pods perpendicular to racemes), slightly drooping(pods show distinct arching habit), or drooping (pods show distinctpointing towards the ground).

Pod (silique) Beak Length: The length of the segment at the end of thepod that does not contain seed (it is a remnant of the stigma and stylefor the flower); visually determined to be short, medium, or long.

Pod (silique) Length: The length of a fully developed pod; visuallydetermined to be short, medium, or long.

Pod (silique) Pedicel Length: The length of the pedicel, ie, the stemthat attaches the pod to the raceme of flowering shoot; visuallydetermined to be short, medium, or long.

Pod (silique) Width: The width of a fully developed pod; visuallydetermined to be narrow, medium, or wide.

Protein Content: The total protein as a percentage by weight of thewhole seed or oil-free meal from mature dried grain; determined by acombustion method (AOCS Official Method Ba 4e-93) or near-infrared (NIR)spectroscopy (AOCS Official Method Am 1-92).

Saturated Fatty Acids: The total of the saturated fatty acids includingC12:0, C14:0, C16:0, C18:0, C20:0, C22:0 and C24:0 as a percentage ofthe oil from unprocessed seed.

Season Type: The growth habit type of canola rated as spring or winter.

Seed Coat Color: The color of the seed coat of a mature seed rated asblack, brown, yellow, mixed, or mottled.

Seed Coat Mucilage: The presence or absence of mucilage in the seedcoat; detected by imbibing seeds with water and monitoring for mucilagethat if present, appears as a halo around the seed. Mucilage is presentin the seed coats of Brassica rapa and absent in the seed coats ofBrassica napus.

Seed Weight: The weight in grams of 1000 seeds at maturity at 5% to 6%moisture.

Seedling Growth Habit: The growth habit of the leaf rosette on youngseedlings, ie, the first 2 to 8 true leaves; rated before flowering asweak (loosely arranged) or strong (closely packed leaves).

Shatter Resistance: The relative amount of plants within a cultivar thatdo not drop seed at maturity; rated as not tested, poor, fair, or good.

Single Gene Converted (Conversion): A cultivar developed by backcrossingor inserting into it by genetic transformation, a single gene or tightlylinked genes, wherein essentially the remainder of the genome isunchanged.

Species: Whether the claimed canola cultivar is a member of the speciesBrassica napus (tetraploid, AACC genomes), Brassica rapa (diploid, AAgenome) or Brassica juncea (tetraploid, AABB genomes).

Stabilized: Reproducibly passed trait from one generation to the nextgeneration of inbred plants of the same cultivar.

Stearic Acid: The total stearic acid (C18:0) in an oil sample obtainedfrom unprocessed seed; measured as a percentage of total oil.

Stem Anthocyanin: The expression of anthocyanin (a purple pigment) inthe stem of plants measured before flowering at the 9- to 11-leaf stagerated as absent or present and, if present, as very weak, weak, medium,strong, or very strong.

Time to Flowering: The number of days from planting to when 50% ofplants of a cultivar show one or more open flowers.

Time to Maturity: The number of days from planting to when all seeds onplants of a cultivar complete filling (about 40% moisture).

White-Rust Resistance: Amount of resistance to white rust (Albugocandida) rated as a percent of susceptible checks; Race 7A measured onBrassica rapa, Races 2V and 7V measured on Brassica napus, and Race 2Ameasured on Brassica juncea.

Winter Hardiness: The relative degree to which plants of a winter-type(fall planted) annual cultivar survives the winter; visually determinedto be poor, fair, good, or excellent.

Yield: The quantity of grain produced in kilograms per hectare (kg/ha),grams per plot (g/plot), or bushels per acre (bu/ac).

Description of G069467

Canola G069467 is an elite doubled-haploid line developed from abi-parental cross, culture of microspores (haploid tissue that normallydevelops into pollen grains) from plants of the cross, chromosomedoubling of the microspores, embryo development and plantletregeneration in culture, seed increase from self pollination, fieldtesting, and selection.

Some of the criteria used to select in various generations include: seedyield, seed protein and oil content, fatty acid composition, lodgingresistance, resistance to glyphosate herbicide, tolerance to diseases,and maturity.

The cultivar G069467 has shown uniformity and stability ofcharacteristics over multiple generations. It has been self pollinatedand increased multiple generations after doubled-haploid plantproduction with careful attention to uniformity of plant type, and nooff-type plants have been found in evaluation.

The present invention relates to a canola plant that expressessubstantially all of the physiological and morphological characteristicsof cultivar G069467. Any plants produced from cultivar G069467 arecontemplated by the present invention and are, therefore, within thescope of this invention. A description of morphological and othercharacteristics of cultivar G069467 is presented in Table 1.

TABLE 1 Morphological and Other Characteristics of Canola Brassica napusG069467 Characteristic Value Season Type (Spring, Winter) Spring Type ofCultivar (Hybrid, Pure Line) Pure Line BEFORE FLOWERING Cotyledon Width(3 = Narrow, 5 = Medium, 7 = Wide) 7 Stem Anthocyanin Intensity (1 =Absent or Very Weak, 3 = Weak, 5 = Medium, 7 = Strong, 3 9 = VeryStrong) Leaf Type (1 = Petiolate, 9 = Lyrate) 1 Leaf Shape (3 = Narrowelliptic, 5 = Wide elliptic, 7 = Orbicular) 3 Leaf Length (3 = Short, 5= Medium, 7 = Long) 7 Leaf Width (3 = Narrow, 5 = Medium, 7 = Wide) 5Leaf Color (at 5-leaf stage) (1 = Light Green, 2 = Medium Green, 3 =Dark Green, 4 = Blue-Green) 3 Leaf Waxiness (1 = Absent or Very Weak, 3= Weak, 5 = Medium, 7 = Strong, 9 = Very Strong) 5 Leaf Texture (1 =Smooth, 9 = Rough) 1 Leaf Lobe Development (1 = Absent or Very Weak, 3 =Weak, 5 = Medium, 7 = Strong, 9 = Very Strong) 5 Leaf Lobe Number(count) 5 Leaf Lobe Shape (1 = Acute, 9 = Rounded) 9 Petiole Length(lobed cultivars only) (3 = Short, 5 = Medium, 7 = Long) 7 Leaf MarginShape (1 = Undulating, 2 = Rounded, 3 = Sharp) 2 Leaf Margin Indentation(observe fully developed upper stem leaves) (1 = Absent or Very 3 Weak(very shallow), 3 = Weak (shallow), 5 = Medium, 7 = Strong (deep), 9 =Very Strong (very deep)) Leaf Attachment to Stem (1 = Complete Clasping,2 = Partial Clasping, 3 = Non-Clasping) 2 AFTER FLOWERING Time toFlowering (days from planting to 50% of plants showing one or more openflowers) 52 Plant Height at Maturity (3 = Short, 5 = Medium, 7 = Tall) 5Plant Growth Habit (1 = Erect, 3 = Semi-Erect, 5 = Intermediate, 7 =Semi-Prostrate, 9 = Prostrate) 1 Flower-Bud Location (1 = Buds abovemost recently opened flowers, 9 = Buds below most 1 recently openedflowers) Petal Color (on first day of flowering) (1 = White, 2 = LightYellow, 3 = Medium Yellow, 4 = Dark 3 Yellow, 5 = Orange, 6 = Other)Petal Length (3 = Short, 5 = Medium, 7 = Long) 5 Petal Width (3 =Narrow, 5 = Medium, 7 = Wide) 5 Petal Spacing (1 = Open, 3 = NotTouching, 5 = Touching, 7 = Slight Overlap, 9 = Strong Overlap) 5 AntherDotting (percentage at opening of flower) (1 = absent, 9 = present(percentage)) 9 Anther Arrangement (observe fully open flower) (1 =Introrse (facing inward), 2 = Erect, 3 3 = Extrorse (facing outward))Anther Fertility (pollen production) (1 = Sterile, 9 = All AnthersShedding) 9 Pod (silique) Length (1 = Short (<7 cm), 5 = Medium (7 to 10cm), 9 = Long (>10 cm)) 1 Pod (silique) Width (3 = Narrow (3 mm), 5 =Medium (4 mm), 7 = Wide (5 mm)) 7 Pod (silique) Angle (1 = Erect, 3 =Semi-Erect, 5 = Horizontal, 7 = Slightly Drooping, 9 = Drooping) 3 Pod(silique) Beak Length (3 = short, 5 = Medium, 7 = Long) 5 Pod PedicelLength (3 = Short, 5 = Medium, 7 = Long) 5 Time to Maturity (days fromplanting to physiological maturity) 108 SEED Seed Coat Color (1 = Black,2 = Brown, 3 = Tan, 4 = Yellow, 5 = Mixed (describe), 6 = Other(specify)) 1 AGRONOMIC Lodging Resistance (1 = fully erect, 3 = 45°angle between stem and the ground, and 5 = flat) 2 REACTION TO DISEASESAND PESTS (0 = Not Tested, 1 = Resistant, 3 = Moderately Resistant, 5 =Moderately Susceptible, 7 = Susceptible, 9 = Hightly Susceptible)Blackleg (Leptosphaeria maculans/Phoma lingam) 1 Fusarium Wilt (Fusariumoxysporum) 1 White Rust (Albugo candida) (list races: (i.e. 2a, 2v, 7a,7v)) 1 REACTION TO CHEMICALS Herbicides Glyphosate Resistance QUALITYOil Content (percentage, whole dry seed basis) 52 Fatty-Acid Composition(percentage of total fatty acids in seed oil) Oleic Acid (C18:1) 74.7Linoleic Acid (C18:2) 14 Linolenic Acid (C18:3) 2.3 Total Saturated Fats6.9 Protein Content (percentage in whole oil-free dry-seed) 42.5Glucosinolates Content (μmoles of total glucosinolates per gram wholeseed, 8.5% moisture) 1 (1 = Very Low (<10 μmol per gram), 2 = Low (10-15μmol per gram), 3 = Medium (15-20 μmol per gram), 4 = High (>20 μmol pergram)) Seed Chlorophyll Content (mg/kg seed, 8.5% moisture) (1 = Low (<8ppm), 2 = Medium (8-15 2 ppm), 3 = High (>15 ppm))

Canola cultivar G069467 has exhibited commercial value in multi-year,multi-location field evaluations. Table 2 shows overall results ofG069467 in the 2009 growing season compared to standard and commercialcheck cultivars in field trials at ten locations in the three seasonzones in western Canada canola production regions. Two locations for theshort-season zone were near Calmar and Ponoka, Alberta; seven locationsfor the medium-season zone were near Neepawa, Manitoba; and Pike Lake,Rosthern, Cudworth, Colonsay, Melfort, and Yorkton, Saskatchewan; andone location for the long-season zone were near Carmen, Manitoba.G069467 demonstrates high grain-oil content composed of high oleic acid,low linolenic acid, low erucic acid, and low total saturated fattyacids. Its grain meal is high in protein and low in glucosinolates. Itsgrain yield is greater than the Western Canada Canola/RapeseedRecommending Committee

(WCC/RRC) standard checks, 46A65 and Q2, and comparable to otherhigh-yielding commercial canola cultivars.

TABLE 2 Mean Grain Yield, Early-Season Vigor, Protein, and OilCharacteristics of Canola Brassica napus G069467 Compared to ControlCultivars Across Field Locations¹ Early- Saturated Grain Season GrainGrain Meal Fatty Oleic Linoleic Linolenic Cultivar Yield Vigor Days toOil Protein Protein Acids C18:1 C18:2 C18:3 Name (kg/ha) (1 to 5)²Maturity (%)^(3,4) (%)^(3,4) (%)^(3,5) (%)⁴ (%)⁴ (%)⁴ (%)⁴ G069467 27782.7 108 52.0 20.4 42.5 6.9 74.7 14.0 2.3 Q2 2490 3.5 109 51.5 20.1 41.57.2 62.2 17.9 10.0 46A65 2427 3.2 106 51.8 21.2 43.9 6.9 64.5 17.5 9.35020 3034 1.9 105 53.3 19.9 42.6 6.9 63.6 17.0 10.8 NX4-105 RR 2675 2.9109 53.6 19.7 42.3 6.7 75.1 13.9 2.1 ¹Ten locations across WesternCanada canola-production zones. ²Scale 1 to 5 where 1 is strong vigorand 5 is weak vigor. ³Percentage of dry matter. ⁴Grain oil, protein, andfatty acids measured by near infrared (NIR) spectroscopy. ⁵Calculatedbased on grain oil and protein measurements.

This invention is also directed to methods for producing a canola plantby crossing a first parent canola plant with a second parent canolaplant, wherein the first or second canola plant is a canola plant fromG069467. Further, both first and second parent canola plants may be fromG069467. Therefore, any methods using G069467 are part of thisinvention: selfing, backcrosses, hybrid breeding, and crosses topopulations. Any plants produced using G069467 as parents are within thescope of this invention.

Useful methods include, but are not limited to, expression vectorsintroduced into plant tissues using a direct gene transfer method suchas microprojectile-mediated delivery, DNA injection, electroporation andthe like. More preferably expression vectors are introduced into planttissues using the microprojectile media delivery with the biolisticdevice Agrobacterium-mediated transformation. Transformant plantsobtained with the protoplasm of the invention are intended to be withinthe scope of this invention.

With the advent of molecular biological techniques that have allowed theisolation and characterization of genes that encode specific proteinproducts, scientists in the field of plant biology developed a stronginterest in engineering the genome of plants to contain and expressforeign genes, or additional, or modified versions of native, orendogenous, genes (perhaps driven by different promoters) in order toalter the traits of a plant in a specific manner. Such foreignadditional and/or modified genes are referred to herein collectively as“transgenes.” Over the last fifteen to twenty years several methods forproducing transgenic plants have been developed, and the presentinvention, in particular embodiments, also relates to transformedversions of the claimed cultivar.

Plant transformation involves the construction of an expression vectorthat will function in plant cells. Such a vector comprises DNAcomprising a gene under control of or operatively linked to a regulatoryelement (for example, a promoter). The expression vector may contain oneor more such operably-linked gene/regulatory element combinations. Thevector(s) may be in the form of a plasmid, and can be used alone or incombination with other plasmids, to provide transformed canola plants,using transformation methods as described below to incorporatetransgenes into the genetic material of the canola plant(s).

Expression Vectors for Canola Transformation: Marker Genes

Expression vectors include at least one genetic marker, operably linkedto a regulatory element (a promoter, for example) that allowstransformed cells containing the marker to be either recovered bynegative selection, i.e., inhibiting growth of cells that do not containthe selectable marker gene, or by positive selection, i.e., screeningfor the product encoded by the genetic marker. Many commonly usedselectable marker genes for plant transformation are well known in thetransformation arts, and include, for example, genes that code forenzymes that metabolically detoxify a selective chemical agent, whichmay be an antibiotic or an herbicide, or genes that encode an alteredtarget that is insensitive to the inhibitor. A few positive selectionmethods are also known in the art.

One commonly used selectable marker gene for plant transformation is theneomycin phosphotransferase II (nptII) gene under the control of plantregulatory signals that confer resistance to kanamycin. Fraley et al.,Proc. Natl. Acad. Sci. U.S.A., 80:4803 (1983). Another commonly usedselectable marker gene is the hygromycin phosphotransferase gene thatconfers resistance to the antibiotic hygromycin. Vanden Elzen et al.,Plant Mol. Biol., 5:299 (1985).

Additional selectable marker genes of bacterial origin that conferresistance to antibiotics include gentamycin acetyl transferase,streptomycin phosphotransferase, aminoglycoside-3′-adenyl transferaseand the bleomycin resistance determinant. Hayford et al., Plant Physiol.86:1216 (1988); Jones et al., Mol. Gen. Genet., 210:86 (1987); Svab etal., Plant Mol. Biol. 14:197 (1990); Hille et al., Plant Mol. Biol.7:171 (1986). Other selectable marker genes confer resistance toherbicides such as glyphosate, glufosinate, or bromoxynil. Comai et al.,Nature 317:741-744 (1985); Gordon-Kamm et al., Plant Cell 2:603-618(1990); and Stalker et al., Science 242:419-423 (1988).

Other selectable marker genes for plant transformation are not ofbacterial origin. These genes include, for example, mouse dihydrofolatereductase, plant 5-enolpyruvylshikimate-3 -phosphate synthase and plantacetolactate synthase. Eichholtz et al., Somatic Cell Mol. Genet. 13:67(1987); Shah et al., Science 233:478 (1986); Charest et al., Plant CellRep. 8:643 (1990).

Another class of marker genes for plant transformation requiresscreening of presumptively transformed plant cells rather than directgenetic selection of transformed cells for resistance to a toxicsubstance, such as an antibiotic. These genes are particularly useful toquantify or visualize the spatial pattern of expression of a gene inspecific tissues and are frequently referred to as reporter genesbecause they can be fused to a gene or gene regulatory sequence for theinvestigation of gene expression. Commonly used genes for screeningpresumptively transformed cells include β-glucuronidase (GUS),β-galactosidase, luciferase and chloramphenicol acetyltransferase.Jefferson, R. A., Plant Mol. Biol. Rep. 5:387 (1987); Teeri et al., EMBOJ. 8:343 (1989); Koncz et al., Proc. Natl. Acad. Sci U.S.A. 84:131(1987); DeBlock et al., EMBO J. 3:1681 (1984).

Recently, in vivo methods for visualizing GUS activity that do notrequire destruction of plant tissue have been made available. MolecularProbes publication 2908, Imagene, Green, T. M., p. 1-4(1993) and Nalewayet al., J. Cell Biol. 115:151a (1991). However, these in vivo methodsfor visualizing GUS activity have not proven useful for recovery oftransformed cells because of low sensitivity, high fluorescentbackgrounds and limitations associated with the use of luciferase genesas selectable markers.

More recently, a gene encoding Green Fluorescent Protein (GFP) has beenutilized as a marker for gene expression in prokaryotic and eukaryoticcells. Chalfie et al., Science 263:802 (1994). GFP and mutants of GFPmay be used as screenable markers.

Expression Vectors for Canola Transformation: Promoters

Genes included in expression vectors must be driven by a nucleotidesequence comprising a regulatory element, for example, a promoter.Several types of promoters are now well known in the transformationarts, as are other regulatory elements that can be used alone or incombination with promoters.

As used herein, “promoter” includes reference to a region of DNAupstream from the start of transcription and involved in recognition andbinding of RNA polymerase and other proteins to initiate transcription.A “plant promoter” is a promoter capable of initiating transcription inplant cells. Examples of promoters under developmental control includepromoters that preferentially initiate transcription in certain tissues,such as leaves, roots, seeds, fibers, xylem vessels, tracheids, orsclerenchyma. Such promoters are referred to as “tissue-preferred.”Promoters that initiate transcription only in certain tissues arereferred to as “tissue-specific.” A “cell type” specific promoterprimarily drives expression in certain cell types in one or more organs,for example, vascular cells in roots or leaves. An “inducible” promoteris a promoter that is under environmental control. Examples ofenvironmental conditions that may affect transcription by induciblepromoters include anaerobic conditions or the presence of light.Tissue-specific, tissue-preferred, cell-type specific, and induciblepromoters constitute the class of “non-constitutive” promoters. A“constitutive” promoter is a promoter that is active under mostenvironmental conditions.

A. Inducible Promoters

An inducible promoter is operably linked to a gene for expression incanola. Optionally, the inducible promoter is operably linked to anucleotide sequence encoding a signal sequence that is operably linkedto a gene for expression in canola. With an inducible promoter, the rateof transcription increases in response to an inducing agent.

Any inducible promoter can be used in the instant invention. See Ward etal., Plant Mol. Biol. 22:361-366 (1993). Exemplary inducible promotersinclude, but are not limited to, that from the ACEI system, whichresponds to copper (Mett et al., PNAS 90:4567-4571 (1993)); In2 genefrom maize, which responds to benzenesulfonamide herbicide safeners(Hershey et al., Mol. Gen Genetics 227:229-237 (1991); and Gatz et al.,Mol. Gen. Genetics 243:32-38 (1994)) or Tet repressor from Tn10 (Gatz etal., Mol. Gen. Genetics 227:229-237 (1991)). A particularly preferredinducible promoter is a promoter that responds to an inducing agent towhich plants do not normally respond. An exemplary inducible promoter isthe inducible promoter from a steroid hormone gene, the transcriptionalactivity of which is induced by a glucocorticosteroid hormone. Schena etal., Proc. Natl. Acad. Sci. U.S.A. 88:0421 (1991).

B. Constitutive Promoters

A constitutive promoter is operably linked to a gene for expression incanola or the constitutive promoter is operably linked to a nucleotidesequence encoding a signal sequence that is operably linked to a genefor expression in canola.

Many different constitutive promoters can be utilized in the instantinvention. Exemplary constitutive promoters include, but are not limitedto, the promoters from plant viruses such as the 35S promoter from CaMV(Odell et al., Nature 313:810-812 (1985)) and the promoters from suchgenes as rice actin (McElroy et al., Plant Cell 2:163-171 (1990));ubiquitin (Christensen et al., Plant Mol. Biol. 12:619-632 (1989) andChristensen et al., Plant Mol. Biol. 18:675-689 (1992)); pEMU (Last etal., Theor. Appl. Genet. 81:581-588 (1991)); MAS (Velten et al., EMBO J.3:2723-2730 (1984)); and maize H3 histone (Lepetit et al., Mol. Gen.Genetics 231:276-285 (1992) and Atanassova et al., Plant Journal 2 (3):291-300 (1992)). The ALS promoter, Xbal/NcoI fragment 5′ to the Brassicanapus ALS3 structural gene (or a nucleotide sequence similarity to saidXbal/NcoI fragment), represents a particularly useful constitutivepromoter. See PCT application WO 96/30530.

C. Tissue-specific or Tissue-preferred Promoters

A tissue-specific promoter is operably linked to a gene for expressionin canola. Optionally, the tissue-specific promoter is operably linkedto a nucleotide sequence encoding a signal sequence that is operablylinked to a gene for expression in canola. Plants transformed with agene of interest operably linked to a tissue-specific promoter producethe protein product of the transgene exclusively, or preferentially, ina specific tissue.

Any tissue-specific or tissue-preferred promoter can be utilized in theinstant invention. Exemplary tissue-specific or tissue-preferredpromoters include, but are not limited to, a root-preferredpromoter—such as that from the phaseolin gene (Murai et al., Science23:476-482 (1983) and Sengupta-Gopalan et al., Proc. Natl. Acad. Sci.U.S.A. 82:3320-3324 (1985)); a leaf-specific and light-induced promotersuch as that from cab or rubisco (Simpson et al., EMBO J.4(11):2723-2729 (1985) and Timko et al., Nature 318:579-582 (1985)); ananther-specific promoter such as that from LAT52 (Twell et al., Mol.Gen. Genetics 217:240-245 (1989)); a pollen-specific promoter such asthat from Zm13 (Guerrero et al., Mol. Gen. Genetics 244:161-168 (1993))or a microspore-preferred promoter such as that from apg (Twell et al.,Sex. Plant Reprod. 6:217-224 (1993)).

Transport of protein produced by transgenes to a subcellular compartmentsuch as the chloroplast, vacuole, peroxisome, glyoxysome, cell wall ormitochondrion or for secretion into the apoplast, is accomplished bymeans of operably linking the nucleotide sequence encoding a signalsequence to the 5′ and/or 3′ region of a gene encoding the protein ofinterest. Targeting sequences at the 5′ and/or 3′ end of the structuralgene may determine, during protein synthesis and processing, where theencoded protein is ultimately compartmentalized.

The presence of a signal sequence directs a polypeptide to either anintracellular organelle or subcellular compartment, or for secretion tothe apoplast. Many signal sequences are known in the art. See, forexample Becker et al., Plant Mol. Biol. 20:49 (1992); Close, P. S.,Master's Thesis, Iowa State University (1993); Knox, C., et al.,“Structure and Organization of Two Divergent Alpha-Amylase Genes fromBarley,” Plant Mol. Biol. 9:3-17 (1987); Lerner et al., Plant Physiol.91:124-129 (1989); Fontes et al., Plant Cell 3:483-496 (1991); Matsuokaet al., Proc. Natl. Acad. Sci. 88:834 (1991); Gould et al., J. Cell.Biol. 108:1657 (1989); Creissen et al., Plant J. 2:129 (1991); Kalderon,et al., A short amino acid sequence able to specify nuclear location,Cell 39:499-509 (1984); Steifel, et al., Expression of a maize cell wallhydroxyproline-rich glycoprotein gene in early leaf and root vasculardifferentiation, Plant Cell 2:785-793 (1990).

Foreign Protein Genes and Agronomic Genes

With transgenic plants according to the present invention, a foreignprotein can be produced in commercial quantities. Thus, techniques forthe selection and propagation of transformed plants, which are wellunderstood in the art, yield a plurality of transgenic plants that areharvested in a conventional manner, and a foreign protein then can beextracted from a tissue of interest or from total biomass. Proteinextraction from plant biomass can be accomplished by known methods,which are discussed, for example, by Heney and Orr, Anal. Biochem.114:92-6 (1981).

According to a particular embodiment, the transgenic plant provided forcommercial production of foreign protein is a canola plant. In anotherpreferred embodiment, the biomass of interest is seed. For therelatively small number of transgenic plants that show higher levels ofexpression, a genetic map can be generated, primarily via conventionalRFLP, PCR and SSR analysis, which identifies the approximate chromosomallocation of the integrated DNA molecule. For exemplary methodologies inthis regard, see Glick and Thompson, Methods in Plant Molecular Biologyand Biotechnology, CRC Press, Boca Raton 269:284 (1993). Map informationconcerning chromosomal location is useful for proprietary protection ofa subject transgenic plant. If unauthorized propagation is undertakenand crosses made with other germplasm, the map of the integration regioncan be compared to similar maps for suspect plants, to determine if thelatter have a common parentage with the subject plant. Map comparisonswould involve hybridizations, RFLP, PCR, SSR and sequencing, all ofwhich are conventional techniques.

Likewise, by means of the present invention, agronomic genes can beexpressed in transformed plants. More particularly, plants can begenetically engineered to express various phenotypes of agronomicinterest. Exemplary genes implicated in this regard include, but are notlimited to, those categorized below:

1. Genes That Confer Resistance to Pests or Disease and That Encode:

A. Plant disease resistance genes. Plant defenses are often activated byspecific interaction between the product of a disease resistance gene(R) in the plant and the product of a corresponding avirulence (Avr)gene in the pathogen. A plant variety can be transformed with clonedresistance genes to engineer plants that are resistant to specificpathogen strains. See, for example, Jones et al., Science 266:789 (1994)(cloning of the tomato Cf-9 gene for resistance to Cladosporium fulvum);Martin et al., Science 262:1432 (1993) (tomato Pto gene for resistanceto Pseudomonas syringae pv. tomato encodes a protein kinase); andMindrinos et al., Cell 78:1089 (1994) (Arabidopsis RSP2 gene forresistance to Pseudomonas syringae).

B. A Bacillus thuringiensis protein, a derivative thereof or a syntheticpolypeptide modeled thereon. See, for example, Geiser et al., Gene48:109 (1986), who disclose the cloning and nucleotide sequence of a Btδ-endotoxin gene. Moreover, DNA molecules encoding δ-endotoxin genes canbe purchased from American Type Culture Collection, Manassas, Va., forexample, under ATCC Accession Nos. 40098, 67136, 31995 and 31998.

C. A lectin. See, for example, the disclosure by Van Damme et al., PlantMolec. Biol. 24:25 (1994), who disclose the nucleotide sequences ofseveral Clivia miniata mannose-binding lectin genes.

D. A vitamin-binding protein such as avidin. See PCT applicationUS93/06487. The application teaches the use of avidin and avidinhomologues as larvicides against insect pests.

E. An enzyme inhibitor, for example, a protease or proteinase inhibitoror an amylase inhibitor. See, for example, Abe et al., J. Biol. Chem.262:16793 (1987) (nucleotide sequence of rice cysteine proteinaseinhibitor); Huub et al., Plant Molec. Biol. 21:985 (1993) (nucleotidesequence of cDNA encoding tobacco proteinase inhibitor I); Sumitani etal., Biosci. Biotech. Biochem. 57:1243 (1993) (nucleotide sequence ofStreptomyces nitrosporeus .alpha.-amylase inhibitor); and U.S. Pat. No.5,494,813 (Hepher and Atkinson, issued Feb. 27, 1996).

F. An insect-specific hormone or pheromone such as an ecdysteroid orjuvenile hormone, a variant thereof, a mimetic based thereon, or anantagonist or agonist thereof. See, for example, the disclosure byHammock et al., Nature 344:458 (1990), of baculovirus expression ofcloned juvenile hormone esterase, an inactivator of juvenile hormone.

G. An insect-specific peptide or neuropeptide which, upon expression,disrupts the physiology of the affected pest. For example, see thedisclosures of Regan, J. Biol. Chem. 269:9 (1994) (expression cloningyields DNA coding for insect diuretic hormone receptor); and Pratt etal., Biochem. Biophys. Res. Comm. 163:1243 (1989) (an allostatin isidentified in Diploptera puntata). See also U.S. Pat. No. 5,266,317 toTomalski et al., who disclose genes encoding insect-specific, paralyticneurotoxins.

H. An insect-specific venom produced in nature by a snake, a wasp, etc.For example, see Pang et al., Gene 116:165 (1992), for disclosure ofheterologous expression in plants of a gene coding for a scorpioninsectotoxic peptide.

I. An enzyme responsible for a hyperaccumulation of a monoterpene, asesquiterpene, a steroid, hydroxamic acid, a phenylpropanoid derivativeor another non-protein molecule with insecticidal activity.

J. An enzyme involved in the modification, including thepost-translational modification, of a biologically active molecule; forexample, a glycolytic enzyme, a proteolytic enzyme, a lipolytic enzyme,a nuclease, a cyclase, a transaminase, an esterase, a hydrolase, aphosphatase, a kinase, a phosphorylase, a polymerase, an elastase, achitinase and a glucanase, whether natural or synthetic. See PCTapplication WO 93/02197 in the name of Scott et al., which discloses thenucleotide sequence of a callase gene. DNA molecules which containchitinase-encoding sequences can be obtained, for example, from the ATCCunder Accession Nos. 39637 and 67152. See also Kramer et al., InsectBiochem. Molec. Biol. 23:691 (1993), who teach the nucleotide sequenceof a cDNA encoding tobacco hornworm chitinase; and Kawalleck et al.,Plant Molec. Biol. 21:673 (1993), who provide the nucleotide sequence ofthe parsley ubi4-2 polyubiquitin gene.

K. A molecule that stimulates signal transduction. For example, see thedisclosure by Botella et al., Plant Molec. Biol. 24:757 (1994), ofnucleotide sequences for mung bean calmodulin cDNA clones; and Griess etal., Plant Physiol. 104:1467 (1994), who provide the nucleotide sequenceof a maize calmodulin cDNA clone.

L. A hydrophobic moment peptide. See PCT application WO 95/16776(disclosure of peptide derivatives of Tachyplesin which inhibit fungalplant pathogens) and PCT application WO 95/18855 (teaches syntheticantimicrobial peptides that confer disease resistance).

M. A membrane permease, a channel former or a channel blocker. Forexample, see the disclosure of Jaynes et al., Plant Sci. 89:43 (1993),of heterologous expression of a cecropin-.beta., lytic peptide analog torender transgenic tobacco plants resistant to Pseudomonas solanacearum.

N. A viral-invasive protein or a complex toxin derived therefrom. Forexample, the accumulation of viral coat proteins in transformed plantcells imparts resistance to viral infection and/or disease developmenteffected by the virus from which the coat protein gene is derived, aswell as by related viruses. See Beachy et al., Ann. rev. Phytopathol.28:451 (1990). Coat protein-mediated resistance has been conferred upontransformed plants against alfalfa mosaic virus, cucumber mosaic virus,tobacco streak virus, potato virus X, potato virus Y, tobacco etchvirus, tobacco rattle virus and tobacco mosaic virus. Id.

O. An insect-specific antibody or an immunotoxin derived therefrom.Thus, an antibody targeted to a critical metabolic function in theinsect gut would inactivate an affected enzyme, killing the insect. Cf.Taylor et al., Abstract #497, Seventh Int'l Symposium on MolecularPlant-Microbe Interactions (Edinburgh, Scotland) (1994) (enzymaticinactivation in transgenic tobacco via production of single-chainantibody fragments).

P. A virus-specific antibody. See, for example, Tavladoraki et al.,Nature 366:469 (1993), who show that transgenic plants expressingrecombinant antibody genes are protected from virus attack.

Q. A developmental-arrestive protein produced in nature by a pathogen ora parasite. Thus, fungal endo α-1,4-D-polygalacturonases facilitatefungal colonization and plant nutrient release by solubilizing plantcell wall homo-α-1,4-D-galacturonase. See Lamb et al., Bio/Technology10:1436 (1992). The cloning and characterization of a gene which encodesa bean endopolygalacturonase-inhibiting protein is described by Toubartet al., Plant J. 2:367 (1992).

R. A developmental-arrestive protein produced in nature by a plant. Forexample, Logemann et al., Bio/Technology 10:305 (1992), have shown thattransgenic plants expressing the barley ribosome-inactivating gene havean increased resistance to fungal disease.

2. Genes That Confer Resistance to an Herbicide:

A. An herbicide that inhibits the growing point or meristem, such as animidazolinone or a sulfonylurea. Exemplary genes in this category codefor mutant ALS and AHAS enzyme as described, for example, by Lee et al.,EMBO J. 7:1241 (1988); and Mild et al., Theor. Appl. Genet. 80:449(1990), respectively.

B. Glyphosate (resistance conferred by, e.g., mutant5-enolpyruvylshikimate-3 -phosphate synthase (EPSPs) genes (via theintroduction of recombinant nucleic acids and/or various forms of invivo mutagenesis of native EPSPs genes), aroA genes and glyphosateacetyl transferase (GAT) genes, respectively), other phosphono compoundssuch as glufosinate (phosphinothricin acetyl transferase (PAT) genesfrom Streptomyces species, including Streptomyces hygroscopicus andStreptomyces viridichromogenes), and pyridinoxy or phenoxy proprionicacids and cyclohexones (ACCase inhibitor-encoding genes), See, forexample, U.S. Pat. No. 4,940,835 to Shah, et al. and U.S. Pat. 6,248,876to Barry et. al., which disclose nucleotide sequences of forms of EPSPswhich can confer glyphosate resistance to a plant. A DNA moleculeencoding a mutant aroA gene can be obtained under ATCC accession number39256, and the nucleotide sequence of the mutant gene is disclosed inU.S. Pat. No. 4,769,061 to Comai. European patent application No. 0 333033 to Kumada et al., and U.S. Pat. No. 4,975,374 to Goodman et al.,disclose nucleotide sequences of glutamine synthetase genes which conferresistance to herbicides such as L-phosphinothricin. The nucleotidesequence of a PAT gene is provided in European application No. 0 242 246to Leemans et al., DeGreef et al., Bio/Technology 7:61 (1989), describethe production of transgenic plants that express chimeric bar genescoding for PAT activity. Exemplary of genes conferring resistance tophenoxy proprionic acids and cyclohexones, such as sethoxydim andhaloxyfop are the Accl-S1, Accl-S2 and Accl-S3 genes described byMarshall et al., Theor. Appl. Genet. 83:435 (1992). GAT genes capable ofconferring glyphosate resistance are described in WO 2005012515 toCastle et. al. Genes conferring resistance to 2,4-D, fop and pyridyloxyauxin herbicides are described in WO 2005107437 and U.S. PatentApplication Serial No. 11/587,893, both assigned to Dow AgroSciencesLLC.

C. An herbicide that inhibits photosynthesis, such as a triazine (psbAand gs+genes) or a benzonitrile (nitrilase gene). Przibila et al., PlantCell 3:169 (1991), describe the transformation of Chlamydomonas withplasmids encoding mutant psbA genes. Nucleotide sequences for nitrilasegenes are disclosed in U.S. Pat. No. 4,810,648 to Stalker, and DNAmolecules containing these genes are available under ATCC Accession Nos.53435, 67441, and 67442. Cloning and expression of DNA coding for aglutathione S-transferase is described by Hayes et al., Biochem. J.285:173 (1992).

3. Genes That Confer or Contribute to a Value-Added Trait, such as:

A. Modified fatty acid metabolism, for example, by transforming a plantwith an antisense gene of stearyl-ACP desaturase to increase stearicacid content of the plant. See Knultzon et al., Proc. Natl. Acad. Sci.U.S.A. 89:2624 (1992).

B. Decreased phytate content—1) Introduction of a phytase-encoding genewould enhance breakdown of phytate, adding more free phosphate to thetransformed plant. For example, see Van Hartingsveldt et al., Gene127:87 (1993), for a disclosure of the nucleotide sequence of anAspergillus niger phytase gene. 2) A gene could be introduced thatreduced phytate content. In maize for example, this could beaccomplished by cloning and then reintroducing DNA associated with thesingle allele which is responsible for maize mutants characterized bylow levels of phytic acid. See Raboy et al., Maydica 35:383 (1990).

C. Modified carbohydrate composition effected, for example, bytransforming plants with a gene coding for an enzyme that alters thebranching pattern of starch. See Shiroza et al., J. Bacteol. 170:810(1988) (nucleotide sequence of Streptococcus mutantsfructosyltransferase gene); Steinmetz et al., Mol. Gen. Genet. 20:220(1985) (nucleotide sequence of Bacillus subtilis levansucrase gene); Penet al., Bio/Technology 10:292 (1992) (production of transgenic plantsthat express Bacillus lichenifonnis α-amylase); Elliot et al., PlantMolec. Biol. 21:515 (1993) (nucleotide sequences of tomato invertasegenes); Sogaard et al., J. Biol. Chem. 268:22480 (1993) (site-directedmutagenesis of barley α-amylase gene); and Fisher et al., Plant Physiol.102:1045 (1993) (maize endosperm starch branching enzyme II).

Methods for Canola Transformation

Numerous methods for plant transformation have been developed includingbiological and physical plant transformation protocols. See, forexample, Mild et al., “Procedures for Introducing Foreign DNA intoPlants” in Methods in Plant Molecular Biology and Biotechnology, GlickB. R. and Thompson, J. E. Eds. (CRC Press, Inc., Boca Raton, 1993) pages67-88. In addition, expression vectors and in vitro culture methods forplant cell or tissue transformation and regeneration of plants areavailable. See, for example, Gruber et al., “Vectors for PlantTransformation” in Methods in Plant Molecular Biology and Biotechnology,Glick B. R. and Thompson, J. E. Eds. (CRC Press, Inc., Boca Raton, 1993)pages 89-119.

A. Agrobacterium-mediated Transformation--One method for introducing anexpression vector into plants is based on the natural transformationsystem of Agrobacterium. See, for example, Horsch et al., Science227:1229 (1985). A. tumefaciens and A. rhizogenes are plant pathogenicsoil bacteria which genetically transform plant cells. The Ti and Riplasmids of A. tumefaciens and A. rhizogenes, respectively, carry genesresponsible for genetic transformation of the plant. See, for example,Kado, C. I., Crit. Rev. Plant Sci. 10:1 (1991). Descriptions ofAgrobacterium vector systems and methods for Agrobacterium-mediated genetransfer are provided by Gruber et al., supra, Mild et al., supra, andMoloney et al., Plant Cell Reports 8:238 (1989). See also, U.S. Pat. No.5,563,055 (Townsend and Thomas), issued Oct. 8, 1996.

B. Direct Gene Transfer—Several methods of plant transformation,collectively referred to as direct gene transfer, have been developed asan alternative to Agrobacterium-mediated transformation. A generallyapplicable method of plant transformation is microprojectile-mediatedtransformation wherein DNA is carried on the surface of microprojectilesmeasuring 1 to 4 μm. The expression vector is introduced into planttissues with a biolistic device that accelerates the microprojectiles tospeeds of 300 to 600 m/s which is sufficient to penetrate plant cellwalls and membranes. Sanford et al., Part. Sci. Technol. 5:27 (1987);Sanford, J. C., Trends Biotech. 6:299 (1988); Klein et al.,Bio/Technology 6:559-563 (1988); Sanford, J. C., Physiol Plant 7:206(1990); Klein et al., Biotechnology 10:268 (1992). See also U.S. Pat.No. 5,015,580 (Christou, et al.), issued May 14, 1991; U.S. Pat. No.5,322,783 (Tomes, et al.), issued Jun. 21, 1994.

Another method for physical delivery of DNA to plants is sonication oftarget cells. Zhang et al., Bio/Technology 9:996 (1991). Alternatively,liposome and spheroplast fusion have been used to introduce expressionvectors into plants. Deshayes et al., EMBO. J., 4:2731 (1985); Christouet al., Proc. Natl. Acad. Sci. U.S.A. 84:3962 (1987). Direct uptake ofDNA into protoplasts using CaCl₂ precipitation, polyvinyl alcohol orpoly-L-ornithine has also been reported. Hain et al., Mol. Gen. Genet.199:161 (1985); and Draper et al., Plant Cell Physiol. 23:451 (1982).Electroporation of protoplasts and whole cells and tissues have alsobeen described. Donn et al., In Abstracts of VIIth InternationalCongress on Plant Cell and Tissue Culture IAPTC, A2-38, p 53 (1990);D'Halluin et al., Plant Cell 4:1495-1505 (1992); and Spencer et al.,Plant Mol. Biol. 24:51-61 (1994).

Following transformation of canola target tissues, expression of theabove-described selectable marker genes allows for preferentialselection of transformed cells, tissues and/or plants, usingregeneration and selection methods now well known in the art.

The foregoing methods for transformation would typically be used forproducing a transgenic variety. The transgenic variety could then becrossed, with another (non-transformed or transformed) variety, in orderto produce a new transgenic variety. Alternatively, a genetic traitwhich has been engineered into a particular canola cultivar using theforegoing transformation techniques could be moved into another cultivarusing traditional backcrossing techniques that are well known in theplant breeding arts. For example, a backcrossing approach could be usedto move an engineered trait from a public, non-elite variety into anelite variety, or from a variety containing a foreign gene in its genomeinto a variety or varieties which do not contain that gene. As usedherein, “crossing” can refer to a simple X by Y cross, or the process ofbackcrossing, depending on the context.

Tissue Culture of Canolas

Further production of the G069467 can occur by self-pollination or bytissue culture and regeneration. Tissue culture of various tissues ofcanola and regeneration of plants therefrom is known. For example, thepropagation of a canola cultivar by tissue culture is described in anyof the following but not limited to any of the following: Chuong et al.,“A Simple Culture Method for Brassica hypocotyls Protoplasts,” PlantCell Reports 4:4-6 (1985); Barsby, T. L., et al., “A Rapid and EfficientAlternative Procedure for the Regeneration of Plants from HypocotylProtoplasts of Brassica napus,” Plant Cell Reports (Spring, 1996);Kartha, K., et al., “In vitro Plant Formation from Stem Explants ofRape,” Physiol. Plant, 31:217-220 (1974); Narasimhulu, S., et al.,“Species Specific Shoot Regeneration Response of Cotyledonary Explantsof Brassicas,” Plant Cell Reports (Spring 1988); Swanson, E.,“Microspore Culture in Brassica,” Methods in Molecular Biology, Vol. 6,Chapter 17, p. 159 (1990).

Further reproduction of the variety can occur by tissue culture andregeneration. Tissue culture of various tissues of soybeans andregeneration of plants therefrom is well known and widely published. Forexample, reference may be had to Komatsuda, T. et al., “Genotype XSucrose Interactions for Somatic Embryogenesis in Soybeans,” Crop Sci.31:333-337 (1991); Stephens, P. A., et al., “Agronomic Evaluation ofTissue-Culture-Derived Soybean Plants,” Theor. Appl. Genet. (1991)82:633-635; Komatsuda, T. et al., “Maturation and Germination of SomaticEmbryos as Affected by Sucrose and Plant Growth Regulators in SoybeansGlycine gracilis Skvortz and Glycine max (L.) Merr.” Plant Cell, Tissueand Organ Culture, 28:103-113 (1992); Dhir, S. et al., “Regeneration ofFertile Plants from Protoplasts of Soybean (Glycine max L. Men.);Genotypic Differences in Culture Response,” Plant Cell Reports (1992)11:285-289; Pandey, P. et al., “Plant Regeneration from Leaf andHypocotyl Explants of Glycine-wightii (W. and A.) VERDC. var.longicauda,” Japan J. Breed. 42:1-5 (1992); and Shetty, K., et al.,“Stimulation of In Vitro Shoot Organogenesis in Glycine max (Merrill.)by Allantoin and Amides,” Plant Science 81:245-251 (1992). Thedisclosures of U.S. Pat. No. 5,024,944 issued Jun. 18, 1991 to Collinset al., and U.S. Pat. No. 5,008,200 issued Apr. 16, 1991 to Ranch etal., are hereby incorporated herein in their entirety by reference.Thus, another aspect of this invention is to provide cells which upongrowth and differentiation produce canola plants having thephysiological and morphological characteristics of canola G069467.

As used herein, the term “tissue culture” indicates a compositioncomprising isolated cells of the same or a different type or acollection of such cells organized into parts of a plant. Exemplarytypes of tissue cultures are protoplasts, calli, plant clumps, and plantcells that can generate tissue culture that are intact in plants orparts of plants, such as embryos, pollen, flowers, seeds, pods, leaves,stems, roots, root tips, anthers, and the like. Means for preparing andmaintaining plant tissue culture are well known in the art. By way ofexample, a tissue culture comprising organs has been used to produceregenerated plants. U.S. Pat. Nos. 5,959,185, 5,973,234 and 5,977,445,describe certain techniques, the disclosures of which are incorporatedherein by reference.

Single-Gene Converted (Conversion) Plants

When the term “canola plant” is used in the context of the presentinvention, this also includes any single-gene conversions of thatvariety. The term “single-gene converted plant” as used herein refers tothose canola plants that are developed by a plant breeding techniquecalled backcrossing, or via genetic engineering, wherein essentially allof the desired morphological and physiological characteristics of avariety are recovered in addition to the single gene transferred intothe variety via the backcrossing technique. Backcrossing methods can beused with the present invention to improve or introduce a characteristicinto the variety. The term “backcrossing” as used herein refers to therepeated crossing of a hybrid progeny back to the recurrent parent,i.e., backcrossing 1, 2, 3, 4, 5, 6, 7, 8 or more times to the recurrentparent. The parental canola plant that contributes the gene for thedesired characteristic is termed the “nonrecurrent” or “donor parent.”This terminology refers to the fact that the nonrecurrent parent is usedone time in the backcross protocol and therefore does not recur. Theparental canola plant to which the gene or genes from the nonrecurrentparent are transferred is known as the recurrent parent as it is usedfor several rounds in the backcrossing protocol (Poehlman & Sleper,1994; Fehr, 1987). In a typical backcross protocol, the original varietyof interest (recurrent parent) is crossed to a second variety(nonrecurrent parent) that carries the single gene of interest to betransferred. The resulting progeny from this cross are then crossedagain to the recurrent parent and the process is repeated until a canolaplant is obtained wherein essentially all of the desired morphologicaland physiological characteristics of the recurrent parent are recoveredin the converted plant, in addition to the single transferred gene fromthe nonrecurrent parent.

The selection of a suitable recurrent parent is an important step for asuccessful backcrossing procedure. The goal of a backcross protocol isto alter or substitute a single trait or characteristic in the originalvariety. To accomplish this, a single gene of the recurrent variety ismodified or substituted with the desired gene from the nonrecurrentparent, while retaining essentially all of the rest of the desiredgenetic, and therefore the desired physiological and morphological,constitution of the original variety. The choice of the particularnonrecurrent parent will depend on the purpose of the backcross. One ofthe major purposes is to add some commercially desirable, agronomicallyimportant trait to the plant. The exact backcrossing protocol willdepend on the characteristic or trait being altered to determine anappropriate testing protocol. Although backcrossing methods aresimplified when the characteristic being transferred is a dominantallele, a recessive allele may also be transferred. In this instance itmay be necessary to introduce a test of the progeny to determine if thedesired characteristic has been successfully transferred.

Many single-gene traits have been identified that are not regularlyselected for in the development of a new variety but that can beimproved by backcrossing techniques. Single-gene traits may or may notbe transgenic, examples of these traits include but are not limited to,male sterility, herbicide resistance, resistance for bacterial, fungal,or viral disease, insect resistance, male fertility, enhancednutritional quality, industrial usage, yield stability and yieldenhancement. These genes are generally inherited through the nucleus.Several of these single-gene traits are described in U.S. Pat. Nos.5,959,185, 5,973,234 and 5,977,445, the disclosures of which arespecifically hereby incorporated by reference.

This invention also is directed to methods for producing a canola plantby crossing a first parent canola plant with a second parent canolaplant wherein the first or second parent canola plant is a canola plantof G069467. Further, both first and second parent canola plants can comefrom the canola G069467. Thus, any such methods using the canolacultivar G069467 are part of this invention: selfing, backcrosses,hybrid production, crosses to populations, and the like. All plantsproduced using canola G069467 as a parent are within the scope of thisinvention, including those developed from varieties derived from canolaG069467. Advantageously, the canola variety could be used in crosseswith other, different, canola plants to produce first generation(F.sub.1) canola hybrid seeds and plants with superior characteristics.The cultivar of the invention can also be used for transformation whereexogenous genes are introduced and expressed by the cultivar of theinvention. Genetic variants created either through traditional breedingmethods using G069467 or through transformation of G069467 by any of anumber of protocols known to those of skill in the art are intended tobe within the scope of this invention.

The invention is also directed to canola meal from seeds of an elitecanola cultivar. In a particular embodiment, the seeds comprise at least44% protein by weight. Canola meal of the present invention can containa characteristic selected from the list consisting of low fiber contentand high protein compared to presently used canola meal.

Oxidative Stability

Stability can be defined as the resistance of a vegetable oil tooxidation and to the resulting deterioration due to the generation ofproducts causing rancidity and decreasing food quality. Tests foroxidative stability attempt to accelerate the normal oxidation processto yield results that can be translated into quality parameters fordifferent food oils and to predict their shelf lives. Stability methodsare also useful to evaluate antioxidants and their effects on protectionof foods against lipid oxidation.

Lipid oxidation in food products develops slowly initially, and thenaccelerates at later stages during storage. The induction period isdefined as the time to reach a constant percentage oxidation of the fatas related to the end of shelf life. The induction period is measuredeither as the time required for a sudden change in rate of oxidation oras the intersection point between the initial and final rates ofoxidation. For vegetable oils containing linoleic and linolenic acid,such as soybean and canola oils, the end-points for acceptability willoccur at relatively low levels of oxidation (peroxide values between 1and 10 Meq/kg).

Factors Affecting Oxidative Stability

The difference in stability between different vegetable oils is due totheir different fatty acid profiles, the effect of processing, initiallevels of oxidation at the start of the storage period, and otherfactors including, minor components, including the presence of metalimpurities, formulation, packaging and environmental storage conditions.From the crude stage to different stages of processing of vegetableoils, some oxidation can take place that will affect the subsequentoxidative stability of the final oil product during storage.

Oxidative Stability Methods

To estimate the oxidative stability of a fat to oxidation, the sample issubjected to an accelerated oxidation test under standardized conditionsand a suitable end-point is chosen to determine the level of oxidativedeterioration. Methods involving elevated temperatures include:

1. Schaal Oven Test

The sample is heated at 50 to 60° C. until it reaches a suitableend-point based on peroxide value or carbonyl value such as theanisidine value. The results of this test correlate best with actualshelf life because the peroxide value end-point of 10 represents arelatively low degree of oxidation. See, limiting peroxide value insection D below.

2. Active Oxygen Method (AOM), Rancimat and Oxidation Stability Index(OSI). See, e.g., U.S. Pat. No. 5,339,294 to Matlock et. al., AOCSMethod 12b-92; and Laubli, M. W. and Bruttel, P. A., JOACS 63:792-795(1986).

Air is bubbled through a sample of oil in special test tubes heated at98-100° C. and the progress of oxidation is followed by peroxide valuedetermination in the AOM test, and by conductivity measurements in theRancimat and OSI tests. The automated Rancimat and OSI tests may be runat temperatures ranging from 100-140° C., and the effluent gases are ledthrough a vessel containing deionized water and the increase inconductivity measured are due to the formation of volatile organic acids(mainly formic acid) by thermal oxidation. The OSI is defined as thetime point in hours of maximum change of the rate of oxidation based onconductivity.

D. Methods to Determine Oxidation - The peroxide value of oils is ameasure of oxidation that is useful for samples that are oxidized torelatively low levels (peroxide values of less than 50), and underconditions sufficiently mild so that the hydroperoxides, which are theprimary products formed by oxidation, are not markedly decomposed. Alimiting peroxide value of 10 meq/kg was specified for refined oils byFAQ/WHO standards (Joint FAQ/WHO Food Standard Program CodexAlimentarius Commission, Report of 16th session of Committee on Fats andOils, London, 1999).

The anisidine test measures high molecular weight saturated andunsaturated carbonyl compounds in oils. The test provides usefulinformation on non-volatile carbonyl compounds formed in oils duringprocessing of oils containing linolenate (soybean and rapeseed). TheTotox value (anisidine value+2 times peroxide value) is used as anempirical measure of the precursor non-volatile carbonyl compoundspresent in processed oils plus any further oxidation products developedafter storage.

While a number of exemplary aspects and embodiments have been discussedabove, those of skill in the art will recognize certain modifications,permutations, additions, and sub-combinations thereof. It is thereforeintended that the following appended claims and claims hereafterintroduced are interpreted to include all such modifications,permutations, additions and sub-combinations as are within their truespirit and scope.

All references, including publications, patents, and patentapplications, cited herein are hereby incorporated by reference to thesame extent as if each reference were individually and specificallyindicated to be incorporated by reference and were set forth in itsentirety herein. The references discussed herein are provided solely fortheir disclosure prior to the filing date of the present application.Nothing herein is to be construed as an admission that the inventors arenot entitled to antedate such disclosure by virtue of prior invention.

DEPOSIT INFORMATION

A deposit of the Dow AgroSciences proprietary canola G069467 disclosedabove and recited in the appended claims has been made with the AmericanType Culture Collection (ATCC), 10801 University Boulevard, Manassas,Va. 20110. The date of deposit was ______. The deposit of ______seedswere taken from the same deposit maintained by ______ since prior to thefiling date of this application. All restrictions upon the deposit havebeen removed, and the deposit is intended to meet all of therequirements of 37 C.F.R. Sections 1.801-1.809. The ATCC accessionnumber is PTA-______. The deposit will be maintained in the depositoryfor a period of 30 years, or 5 years after the last request, or for theeffective life of the patent, whichever is longer, and will be replacedas necessary during that period.

What is claimed is:
 1. A seed of canola designated G069467, wherein arepresentative sample of seed of said cultivar was deposited under ATCCAccession No. PTA-______.
 2. A canola plant, or a part thereof, producedby growing the seed of claim
 1. 3. A method of introducing a desiredtrait into canola G069467, wherein the method comprises: (a) crossing aG069467 plant, wherein a representative sample of seed was depositedunder ATCC Accession No. PTA-______, with a plant of another canolacultivar that comprises a desired trait to produce F₁ progeny plants,wherein the desired trait is selected from the group consisting of malesterility, herbicide resistance, insect resistance, and resistance tobacterial disease, fungal disease or viral disease; (b) selecting one ormore progeny plants that have the desired trait to produce selectedprogeny plants; (c) crossing the selected progeny plants with theG069467 plants to produce backcross progeny plants; (d) selecting forbackcross progeny plants that have the desired trait and physiologicaland morphological characteristics of canola G069467 to produce selectedbackcross progeny plants; and (e) repeating steps (c) and (d) three ormore times to produce selected fourth or higher backcross progeny plantsthat comprise the desired trait and an oleic acid value of about 70% andan α-linolenic acid value of less than about 3%.
 4. The method of claim3, wherein the plants further comprise a yield greater than about 2100kg/ha, a protein value of greater than 44%, or a glucosinolate value ofless than 12%.
 5. The method of claim 3, wherein the plants furthercomprise resistance to Blackleg (Leptosphaeria maculans), Fusarium wilt,or White Rust.
 6. The method of claim 3, wherein the plants furthercomprise herbicide resistance to an herbicide selected from the groupconsisting of imidazolinone, sulfonylurea, glyphosate, glufosinate,L-phosphinothricin, triazine, Clearfield, Dicamaba, 2,4-D, andbenzonitrile.
 7. The method of claim 3, wherein the plants comprise allof the physiological and morphological characteristics of canola G069467as shown in Tables 1 and
 2. 8. A canola plant produced by the method ofclaim 3, wherein the plant has the desired trait and desired traitcomprises an oleic acid value of about 70% and an α-linolenic acid valueof less than about 3%.
 9. The canola plant of claim 8, wherein thedesired trait further comprises herbicide resistance and the resistanceis conferred to an herbicide selected from the group consisting ofimidazolinone, sulfonylurea, glyphosate, glufosinate,L-phosphinothricin, triazine, Clearfield, Dicamaba, 2,4-D, andbenzonitrile.
 10. The canola plant of claim 8, wherein the desired traitfurther comprises insect resistance and the insect resistance isconferred by a transgene encoding a Bacillus thuringiensis endotoxin.11. The canola plant of claim 8, wherein the desired trait furthercomprises resistance to Blackleg, Fusarium wilt, or White Rust.
 12. Thecanola plant of claim 8, wherein the plant comprises all of thephysiological and morphological characteristics of canola G069467, asshown in Tables 1 and
 2. 13. A method of modifying fatty acid metabolismor modifying carbohydrate metabolism of canola G069467 wherein themethod comprises: (a) crossing a G069467 plant, wherein a representativesample of seed was deposited under ATCC Accession No. PTA-______, with aplant of another canola cultivar to produce F₁ progeny plants thatcomprise a nucleic acid molecule encoding an enzyme selected from thegroup consisting of phytase, fructosyltransferase, levansucrase,alpha-amylase, invertase and starch branching enzyme or encoding anantisense of stearyl-ACP desaturase; (b) selecting one or more progenyplants that have said nucleic acid molecule to produce selected progenyplants; (c) crossing the selected progeny plants with the G069467 plantsto produce backcross progeny plants; (d) selecting for backcross progenyplants that have said nucleic acid molecule and physiological andmorphological characteristics of canola G069467 to produce selectedbackcross progeny plants; and (e) repeating steps (c) and (d) three ormore times to produce selected fourth or higher backcross progeny plantsthat comprise said nucleic acid molecule and have an oleic acid value ofabout 70% and an α-linolenic acid value of less than about 3%.
 14. Themethod of claim 13, wherein the plants further comprise a yield greaterthan about 2100 kg/ha, a protein value of greater than 44%, or aglucosinolate value of less than 12%.
 15. The method of claim 13,wherein the plants further comprise resistance to Blackleg(Leptosphaeria maculans), Fusarium wilt, or White Rust.
 16. The methodof claim 13, wherein the plants further comprise herbicide resistance toan herbicide selected from the group consisting of imidazolinone,sulfonylurea, glyphosate, glufosinate, L-phosphinothricin, triazine,Clearfield, Dicamaba, 2,4-D, and benzonitrile.
 17. The method of claim13, wherein the plants comprise all of the physiological andmorphological characteristics of canola G069467 as shown in Tables 1 and2.
 18. A canola plant produced by the method of claim 13, wherein theplant comprises the nucleic acid molecule and has an oleic acid value ofabout 70% and an α-linolenic acid value of less than about 3%.
 19. Acanola plant produced by the method of claim 13, wherein the plantsfurther comprise a yield greater than about 2100 kg/ha, a protein valueof greater than 44%, or a glucosinolate value of less than 12%.
 20. Acanola plant produced by the method of claim 13, wherein the plantscomprise all of the physiological and morphological characteristics ofcanola G069467 as shown in Tables land 2.